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The IAC template was consistently amplified in all samples as illustrated by a Ct value of approximately 27 ruling out a PCR inhibition in the sample preparations.
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Using these optimizations, single sequence reads of 750 1000 bp from single regular and XL-length double-stranded PCR-derived DNA templates were consistently obtained, without the need for subcloning or large-insert libraries.
Pol I transcription from the nucleosomal template (N) was consistently 10-fold less than that from the same amount of nucleosome-free DNA template (F), suggesting that the nucleosome presents a significant but not insurmountable barrier to efficient transcription elongation by Pol I. We next asked whether Pol I could transcribe through a template with an array of nucleosomes.
Three 'no template' controls were consistently negative for each reaction.
In particular, the melting curve from 1 ng of mutant template was sufficiently different from wild-type to identify the mutation, and this distinct melting profile was consistently seen across all the other template amounts investigated.
The α-solenoid fold of β-karyopherins such as Importin-β was consistently identified as a high-confidence template for CENP-I structural modeling.
The EP4 receptor was consistently the most abundant receptor transcript (shortest number of cycles to exponential amplification in identical template, ANOVA p = 0.0002, Figure 1b).
It was consistently funny.
It was consistently, wonderfully odd.
The concert was consistently impressive.
It was consistently good and consistently inexpensive.
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